Question: Differential binding analysis
0
gravatar for marina-orlova
6.1 years ago by
Russian Federation
marina-orlova80 wrote:

Hello

Could you please tell me if there is any way to make differential binding analysis with chip-chip and chip-seq data (one sample is  from chip-chip experiment (.cel files and gff3 file with peaks) and the other is .bam from ChiP-seq (.bam files and gff3 with peaks)).

chip-on-chip chip-seq diffbind • 3.0k views
ADD COMMENTlink modified 5.5 years ago by vivekbhr630 • written 6.1 years ago by marina-orlova80

I have no experience with chip-on-chip, but comparing two sample (and two conditions?) with one technique is already difficult to interpret as you cannot know to what extent the differences you see are due to the conditions or to the within condition variability. Comparing two samples from two techniques is even worse...!

ADD REPLYlink written 6.1 years ago by dariober11k
1
gravatar for seidel
6.1 years ago by
seidel7.1k
United States
seidel7.1k wrote:

There's no robust way that I'm aware of. As dariober hinted at, there are too many caveats to consider when comparing peaks across two very different technologies. However, depending on your ultimate objective, you could perform an ad hoc analysis that could tell you a few things. You could use bedtools or the IRanges capabilities of R to tell you how well the peaks overlap between the two experiments - that's the most basic level of differential analysis. You could then use those same tools to define the common regions called as peaks between both platforms, and compare the distributions of scores (i.e. each might be a ratio over background) under the common areas to see how comparable they are. To quantify differences you might have to normalize them to each other, or transform them to a similar scale (i.e. z-score, etc.), or examine them by rank - a lot depends on what the data looks like, and what you're really trying to get out of it.

ADD COMMENTlink written 6.1 years ago by seidel7.1k

Thank you a lot for your answer, could you please advise how I can normalize chip to input ratio to compare them to each other (please advise what to do in general terms)? The first image is peaks of ChiP-seq and the other is peaks of ChiP-ChiP

ADD REPLYlink modified 6.1 years ago • written 6.1 years ago by marina-orlova80
1
gravatar for vivekbhr
5.5 years ago by
vivekbhr630
Germany
vivekbhr630 wrote:

To compare two conditions in ChIP-Seq, you can use the DiffBind package in Bioconductor ..

http://www.bioconductor.org/packages/release/bioc/html/DiffBind.html

ADD COMMENTlink written 5.5 years ago by vivekbhr630
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